Bothrops alternatus snake venom induces apoptosis of skeletal muscle cells in a rabbit model / O veneno da serpente Bothrops alternatus induz apoptose nas células musculares de coelhos

Este estudo objetiva reportar a ocorrência de apoptose in vivo induzida pelo veneno da serpente Bothrops alternatus em células musculares esqueléticas. Cinco coelhos machos, adultos, receberam 150µg/kg de veneno no músculo vasto lateral, enquanto outros cinco animais receberam 0,1% de BSA diluído em PBS no mesmo local. Após 12 horas, os animais foram eutanasiados, e amostras do local de inoculação foram coletadas para análise histopatológica. Foram evidenciadas necrose e hemorragia nas células musculares. Além disso, a análise imuno-histoquímica para identificação de caspase-3 ativada revelou marcações granulares e agregadas no citoplasma das células musculares, compatíveis com o processo de apoptose. Este é o primeiro relato que confirma o veneno de B. alternatus como causador de apoptose in vivo em células musculares esqueléticas.(AU)

Use of EDTA in the treatment of local tissue damage caused by the Bothrops alternatus venom / Utilização do EDTA no tratamento da lesão tecidual local causado pelo veneno de Bothrops alternatus

Twelve adult rabbits were distributed in three groups and received on the femoral biceps region, via intradermal injection (ID), 25µg of Bothrops alternatus venom dissolved in NaCl 0.9% and diluted in 0.25mL of phosphate buffered saline (PBS). Thirty minutes later, the group G1 received 0.25mL of phosphate buffered saline (PBS) ID while to G2 and G3 25mg of ethylenediamine tetraacetic acid (EDTA) dissolved in 0.25mL of PBS were administered via intramuscular (IM) and intravenous (IV) injection, respectively. Evaluations included local lesion and blood profile of all animals, before (time zero) and at 1, 2, 3, 4, 5, 6, 12, 18 and 24h after venom administration. All animal treated with PBS (G1) and EDTA IV (G3) presented increase of nociceptive stimuli at the site of inoculation of the venom, followed by moderate edema that persisted for 24h. Animals treated with IM EDTA (G2) only manifested increase of nociceptive stimuli at the site of injection 1h after treatment with discrete local edema between 12 and 24h. In relation to the local hemorrhagic halo no differences were found amongst the studied groups. Blood profile revealed significant decrease of segmented neutrophils in all groups. There was also increase in triglycerides and decrease in total protein and albumin in all groups. The local lesion was not altered by the treatments.(AU)

Doze coelhos adultos, distribuídos em três grupos, receberam, na região de bíceps femoral, por via intradérmica (ID), 25µg de veneno de Bothrops alternatus, dissolvidos em NaCl 09%, diluído em 0,25mL de tampão salina fosfato (PBS). Trinta minutos após o desafio, o grupo G1 recebeu 0,25mL de (PBS) ID, e os grupos G2 e G3 receberam 25mg de ácido etilenodiamino tetra-acético (EDTA), dissolvidos em mL de PBS por via intramuscular (IM) e intravenosa (IV), respectivamente. Foram avaliados lesão local e perfil sanguíneo de todos os animais, antes - tempo zero, e à uma, às duas, três, quatro, cinco, seis, 12, 18 e 24 horas após a injeção do veneno. Tanto os animais tratados com PBS (G1) como os animais tratados com EDTA IV (G3) apresentaram aumento do estímulo nociceptivo no local da administração do veneno, seguido por moderado edema, que perdurou por 24h. Os animais tratados com EDTA IM (G2) somente manifestaram aumento do estímulo nociceptivo local uma hora após tratamento e discreto edema local entre 12 e 24 horas. Em relação ao halo hemorrágico, não houve diferença entre os três grupos estudados. No perfil hematológico, observou-se diminuição significativa dos neutrófilos segmentados nos três grupos estudados. Da mesma forma, houve aumento dos triglicerídeos e diminuição da proteína total e albumina em todos os grupos. Conclui-se que a lesão local não foi alterada pelos tratamentos.(AU)

Perfil hematológico de éguas Quarto de Milha alimentadas com feno ou haylage de Tifton-85 ( Cynodon spp. ) / Hematological panel of Quarter Horse mares fed Tifton 85 (Cynodon spp. ) hay and haylage

RESUMO O capim Tifton-85 (Cynodon spp.) vem sendo utilizado na alimentação de equinos na forma de pré-secado (haylage) e feno, todavia não existem estudos conclusivos a respeito da influência da haylage sobre o estado de saúde dessa espécie. Portanto, esta pesquisa objetivou avaliar o perfil sanguíneo como indicador de higidez de éguas Quarto de Milha alimentadas com haylage e feno de Tifton-85. A gramínea utilizada foi cortada com 30 dias de crescimento. Para produção da haylage, a planta cortada permaneceu no campo até atingir 70% de matéria seca, quando foi colhida. Foram utilizadas 12 éguas Quarto de Milha, com idade entre oito e 12 anos e com peso vivo médio inicial de 451,6kg. Os animais foram distribuídos aleatoriamente em dois grupos (n=6), sendo o grupo 1 alimentado com feno e o grupo 2 alimentado com haylage, por um período de 28 dias. Foram realizadas cinco coletas de sangue em intervalos de uma semana, para a determinação do perfil hematológico. Em relação ao eritrograma, observou-se discreta diminuição da concentração de hemoglobina, do hematócrito e do volume globular médio (VGM) em ambos os grupos. No leucograma, houve aumento dos leucócitos totais nas coletas iniciais em ambos os grupos, mas sem significado clínico. Conclui-se que éguas adultas Quarto de Milha, após a ingestão de feno e haylage de Tifton-85, não apresentaram alterações no perfil hematológico que indicassem prejuízos à saúde.(AU)

ABSTRACT Tifton-85 grass (Cynodon spp) has been used in equine nutrition in the form of pre-dried (haylage) and hay, however, there are no conclusive studies about the influence of haylage on blood profile of this species. Therefore, the aim of this study was to evaluate the blood profile as health indicators of Quarter Horse mares were fed haylage and Tifton-85 hay. The used grass was cut at 30 days of growth. For the production of haylage, this plant remained cut in the field until it had 70% of dry matter, when it was collected. Twelve Quarter Horse mares, aged 8-12 years, with average weight of 451.58kg were used. The animals were distributed randomly into two groups (n= 6), group 1 fed hay and group 2 fed haylage, both for 28 days. Blood samples were collected at one week intervals each for determining the hematological parameters. In the erythrogram, a decrease in the hemoglobin concentration, packed cell volume and mean cell volume after the administration in both groups were detected. The white blood count increased in both groups, but the values remained within normal limits without clinical significance. In conclusion, adult Quarter Horse mares fed Tifton-85 hay and haylage showed no alterations in hematology.(AU)

Effects of different extracts of the mushroom Agaricus blazei Murill on the hematologic profile of mice with Ehrlich tumor / Efeitos de diferentes extratos do cogumelo Agaricus blazei Murill sobre o perfil hematológico de camundongos com tumor de Ehrlich

The aim of this study was to investigate the effect of the mushroom Agaricus blazeii Murril (ABM) extracts on the hematological profile of Swiss mice bearing an Ehrlich solid tumor. Three fractions (total extract, polysaccharides, and supernatant) of ABM extracts obtained by four methods (ultrasonic or water bath, at pH 4 or pH 7) were administered to mice over 21 days. Polysaccharide solutions were analyzed by gas and liquid chromatography that showed both mannose and glucose concentrations. The method of extraction influenced the degree of glucose polymerization and the mannose/glucose relationship. The treatment with ABM supernatant at pH 7 and water bath was associated with reduced concentrations of leukocytes and lymphocytes and altered the percentage of CD4+ and CD8+ lymphocytes in Ehrlich tumor-bearing mice. The treatment with the ABM extract in water bath and ultrasound at pH 4 resulted in lower lymphocyte counts, regardless of tumor presence, and greater granulocyte values in mice with Ehrlich tumor than in controls. We concluded that different fractions and methods of extraction of A. blazei produced differing blood profiles in mice inoculated with the Ehrlich tumor.

O objetivo deste estudo foi investigar o efeito de diferentes extratos do cogumelo Agaricus blazeii Murril (ABM) sobre o perfil hematológico de camundongos Swiss portadores de tumor de Ehrlich sólido. Três frações (extrato total, polissacarídeos e sobrenadante) dos extratos de ABM foram obtidas por quatro métodos (sonificador, banho-maria, em pH 4 ou pH 7) e administradas para camundongos durante 21 dias. Soluções de polissacarídeos foram analisadas por cromatografia gasosa e líquida, que mostraram concentrações de glucose e manose. O método de extração influenciou o grau de polimerização da glicose e a relação manose/glucose. O tratamento com o sobrenadante de ABM (em pH 7 e banho-maria) estava associado com reduzidas concentrações de leucócitos e linfócitos, além de alterar a porcentagem de linfócitos CD4+ e CD8+ em camundongos portadores de tumor sólido de Ehrlich. O tratamento com extratos de ABM, obtidos tanto em banho-maria como no sonificador em pH 4, resultou nas mais baixas contagens de linfócitos, independentemente da presença do tumor, e nos maiores valores de granulócitos em camundongos com tumor de Ehrlich. Conclui-se que os diferentes métodos de extração com as respectivas frações de A. blazei são capazes de intereferir no perfil hematológico de camundongos com tumor sólido de Ehrlich.

Pathological effects of Cryptostegia venusta toxicity in goats and rats.

Here, we aimed to determine the toxicity of Cryptostegia venusta in goats and rats. We orally administered a single 60 g dose of shredded C. venusta leaves per kilogram of body weight to three goats. The animals were necropsied after death, and tissue sections were collected and routinely processed for histopathological analyses. Additionally, we separated 25 adult male Wistar rats (each weighing about 150 g) into five groups: an untreated control group and groups orally treated with 1, 3, 10, or 60 g/kg doses. Rats were sacrificed 72 h after administration of the C. venusta extract, and tissue sections collected for histopathological analyses. All goats presented signs of apathy, salivation, frequent urination, and eventually fatigue 4-6h after receiving C. venusta. Two goats died 20 h after administration, and the third was sacrificed in extremis. The only histopathological finding observed in the goats was lung edema. No rats died during the experimental period or presented any clinical signs or macroscopic lesions. However, both goats and rats exhibited degeneration and multifocal necrosis of cardiac muscle fibers. From our results, we conclude that the C. venusta plant is capable of promoting cardiotoxicity.

Eritrograma e variantes de hemoglobina em caprinos da raça Canindé / Erythrogram and hemoglobin types in Canindé goats

The erythrogram and hemoglobin types in Canindé goats were determined. Twenty blood samples were collected from the animals and used for determination of packed cell volume (PCV), erythrocytes counts, hemoglobin concentration, calculation of mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), and electrophoretic separation of hemoglobin types. The values for erythrogram were: PCV 34.6±4.62 percent, hemoglobin 10.1±1.05mg/dl, erythrocytes 15.3±4.54x106/mm3, MCV 24.0±5.83fl, and MCHC 29.3±2.44g/dl. Electrophoretic separation of hemoglobin types revealed only one type characterized as hemoglobin A (HbAA).

Conditioning taste aversion to Mascagnia rigida (Malpighiaceae) in sheep.

The objective of this study was to determine if sheep could be averted to Mascagnia rigida, a toxic plant found in the semiarid region of northeastern Brazil. Twelve female sheep naïve to M. rigida were randomly allocated to two treatment groups: control (treated with 15 mL water orally by a drenching gun) and lithium group (treated with 150 mg LiCl/kg body weight orally by a drenching gun). For conditioning, sheep were allowed to feed on M. rigida leaves for 15 min, followed by LiCl or water administration. The time spent eating M. rigida leaves was measured. The conditioning was repeated daily until the LiCl-treated sheep stopped eating M. rigida, which occurred at days 2 and 3. Persistence trials were conducted on day 10, 24, 40, 55, and 70 of the trial using single-choice tests. There was no difference between the two treatment groups with respect to the consumption of M. rigida on the first day of aversion conditioning. On the second day, three out of the six sheep in the lithium group did not eat the leaves, but on the third day, all the sheep in the lithium group did not ingest M. rigida. This aversion persisted throughout all the persistence trials. This indicates that sheep can be easily conditioned by using lithium chloride to avoid eating M. rigida.

Toxicity in goats caused by oleander (Nerium oleander).

Cases of poisoning by oleander (Nerium oleander) were observed in several species, except in goats. This study aimed to evaluate the pathological effects of oleander in goats. The experimental design used three goats per group: the control group, which did not receive oleander and the experimental group, which received leaves of oleander (50 mg/kg/day) for six consecutive days. On the seventh day, goats received 110 mg/kg of oleander leaves four times at one-hourly interval. A last dose of 330 mg/kg of oleander leaves was given subsequently. After the last dose was administered, clinical signs such as apathy, colic, vocalizations, hyperpnea, polyuria, and moderate rumen distention were observed. Electrocardiogram revealed second-degree atrioventricular block. Death occurred on an average at 92 min after the last dosing. Microscopic evaluation revealed renal necrosis at convoluted and collector tubules and slight myocardial degeneration was observed by unequal staining of cardiomyocytes. Data suggest that goats appear to respond to oleander poisoning in a manner similar to other species.

Comparative effects of prolonged administration of cyanide, thiocyanate and chokecherry (Prunus virginiana) to goats.

The aim of the present study was to determine and compare the clinical, hematological, biochemical and histopathological changes induced by cyanide, thiocyanate and chokecherry (Prunus virginiana) in goats. Sixteen Boer-Spanish cross-bred female goats were divided into four treatment groups: (1) control, (2) potassium cyanide (KCN) at 3.8 mg kg(-1) day(-1), (3) potassium thiocyanate (KSCN) at 4.5 mg kg(-1) day(-1) and (4) ground frozen chokecherry leaves and flowers at a target dose of 2.5 mg HCN kg(-1) day(-1), all for 4 weeks. Clinical signs were observed in two goats treated with chokecherry. Only sporadic changes were found in the hematological and blood chemical panel. Goats treated with chokecherry and thiocyanate had an increased number of vacuoles in the colloid of thyroid glands. Spongiosis and spheroids were found in the mesencephalon from goats treated with KCN and chokecherry. These findings suggest the thyroid lesions can be attributed to thiocyanate, whereas the effects on the nervous system were most likely caused by cyanide.

Determinação das atividades séricas de creatina quinase, lactato desidrogenase e aspartato aminotransferase em eqüinos de diferentes categorias de atividade / Determination of serum activities of creatine kinase, lactate dehydrogenase, and aspartate aminotransferase in horses of different activities classes

The creatine kinase (CK), lactate dehydrogenase (LDH), and aspartate aminotransferase (AST) seric activities in horses of different activity classes (athlete, traction, and reproduction), were compared. Fifty-eight horses were alloted into three groups - group 1 with 20 athletes, "vaquejada" competitors; group 2 with 20 breeding horses; and group 3 with 18 draft horses, averaging 10 working hours daily. The average values for CK serum activity were 80.2, 83.9, and 94.4 U/l in groups 1, 2, and 3, respectively. Result of group 3 was significantly different from the other groups. The averages values for LDH were 102.5, 98.6, and 112.8 U/l in groups 1, 2, and 3, respectively, with no statistical difference between groups. The AST averages were 56.8, 33.0, and 50.1 U/l in groups 1, 2, and 3, respectively, with group 2 significantly differing from the others. Clinical biochemistry values of muscular function in horses varied according to activity category.

Effects of long-term cyanide ingestion by pigs.

Animal performance and health status are adversely affected by long-term cyanide ingestion; however, the effects of cyanide ingestion by pigs have not been fully determined. The aim of the present study was to determine the effects of prolonged exposure to different doses of potassium cyanide (KCN) in growing-finishing swine. Twenty-four pigs, 45 days of age, were divided into four equal groups and treated with different doses of KCN: 0, 2.0, 4.0 or 6.0 mg per kg body weight per day for 70 consecutive days. The results showed a significant alteration in thiocyanate, creatinine and urea levels and in alanine aminotransferase activity of swine dosed with 4.0 and 6.0 mg/kg/KCN. Thyroid weight was significantly increased in those pigs from 4.0 mg/kg KCN group, but no change in cholesterol, triiodothyronine or thyroline levels were observed. Body and carcase weights, body weight gain, and bacon thickness were not affected by KCN treatment. The histopathological study revealed increased numbers of vacuoles in the colloid of thyroid follicles, degeneration of cerebellar white matter and Purkinje cells, degeneration of renal tubular epithelial cells, caryolysis and pyknosis in hepatocytes, and disturbance of the normal lobular architecture of the liver in all treated pigs. Thus, long-term administration of KCN to swine affects several tissues and could adversely affect animal production.

Avaliação do fluido peritoneal de asininos / Evaluation of peritoneal fluid from asinines

The peritoneal fluid from six female health donkeys was evaluated. In each animal, it was carried out eight abdominocentesis with 40x12 needles, at 72-hour intervals. All animals presented clinical changes, and there was higher frequency of yellow-tinged fluid with turbidity aspect. Means, and respective standard deviations, for cytology and protein concentration were: erythrocytes, 24.7±11.7x10³/mm³; leukocytes, 15.8±6.79x10³/mm³; mesothelial cells, 3.85±1.30x10³/mm³; neutrophils, 10.9±4.51x10³/mm³; lymphocytes, 1.41±0.51x10³/mm³; macrophages, 0.98±0.55x10³/mm³; eosinophils, 2.48±1.36x10³/mm³, and total proteins, 1.97±0.31 g/dl.

Acute cattle intoxication from Nerium oleander pods.

Seven outbreaks of acute intoxication from oleander (Nerium oleander) in cattle were reported in Northeast of Brazil. A total of 92 cattle were poisoned by oleander in 7 different herds; 57 animals died (67% of affected cattle). All cases reported here occurred during dry season. Two of the outbreaks resulted from offering oleander triturated and mixed with fodder. In the other cases, accidental ingestion of residual parts of oleander derived from pruning or cutting plants on grazing land were responsible. Clinical signs were diverse; the most common were locomotion disturbances, diarrhoea, depression and sudden death. Postmortem findings varied from no significant lesions to widespread haemorrhage. The presence of oleander leaves in the rumen was noted in all cases. The lack of information about the toxicity of oleanders was the main cause for the accident, which prompt us to stimulate wide divulgation of the common toxic plants.

Clinical and pathological effects of short-term cyanide repeated dosing to goats.

The purpose of this work is to determine and describe the effects of subacute cyanide toxicity to goats. Eight female goats were divided into two groups. The first group of five animals was treated with 8.0 mg KCN kg(-1) body weight day(-1) for seven consecutive days. The second group of three animals was treated with water as controls. Complete physical examination, including observation for behavior changes, was conducted before and after dosing. One treated animal was euthanized immediately after dosing. Later, two of the remaining treated animals and a control goat were euthanized after a 30-day recovery period. Euthanized animals were necropsied and tissues were collected and prepared for histologic studies. Clinical signs in treated goats were transient and included depression and lethargy, mild hyperpnea and hyperthermia, arrhythmias, abundant salivation, vocalizations, expiratory dyspnea, jerky movements and head pressing. Two goats developed convulsions after day 3 of treatment. One animal developed more permanent behavioral changes as she became less dominant and aggressive. Histologic changes included mild hepatocellular vacuolation and degeneration, mild vacuolation and swelling of the proximal convoluted tubules of the kidneys and spongiosis of the white matter (status spongiosis) of the cerebral white tracts, internal capsule, cerebellar peduncles, spinal cord and peripheral nerves. In summary, sub-lethal cyanide intoxication in goats resulted in behavioral changes, and during the treatment period animals showed delayed signs of toxicity. Significant histologic lesions in goats were observed and need to be characterized further.

Neuropathologic study of long term cyanide administration to goats.

Cyanogenic glycosides, which release cyanide, are present in several plant species of high importance for animal production, such as cassava and sorghum. Several human neurological diseases have been associated with chronic cyanide exposure. On the other hand, these effects in ruminants are almost unknown. Thus, the objective of the present study was to determine the long-term lesions of the central nervous system (CNS) caused by daily administration of potassium cyanide (KCN) to goats. Thirty-four male goats were divided into five groups, respectively treated orally with 0 (control), 0.3, 0.6, 1.2 or 3.0 mg KCN/kg/day for 5 months. At the end of the experiment, the whole CNS of each animal was collected for histopathology and immunohistochemistry for apoptotic markers (BAX, BCl2 and CPP32) and for glial fibrillary acid protein (GFAP; vimentin). The results showed the presence of spheroids in the pons, medulla oblongata, and ventral horn of the spinal cord, gliosis and spongiosis in medulla oblongata, gliosis in the pons, and damaged Purkinje cells in the cerebellum from goats that received the higher cyanide dose. In goats from the 1.2 mg KCN/kg group we observed congestion and hemorrhage in the cerebellum, and spheroids in the spinal cord. Gliosis was confirmed by GFAP protein expression. Immunohistochemistry for apoptotic markers and typical apoptotic morphology suggested apoptosis did not participate in the pathogenesis of the observed lesions. Thus, chronic cyanide exposure can promote neuropathological lesions also in goats, and this species can be a useful ruminant model to study the neurotoxic effects of long-term cyanide exposure.

Physiopathological effects of the administration of chronic cyanide to growing goats--a model for ingestion of cyanogenic plants.

Ingestion of cyanogenic plants, such as cassava and sorghum, has been associated with goitre and tropical pancreatic diabetes in both humans and animals. Thus, the objective of the present study was to determine the toxic effects on the thyroid and pancreas in growing goats of prolonged exposure to potassium cyanide (KCN). Thirty-four male goats were divided into five groups dosed with KCN at 0 (control). 0.3, 0.6, 1.2 or 3.0 mg/kg daily for 5 months. Blood samples were obtained in order to determine the glucose, cholesterol, thyroxine (T4), triiodothyronine (T3) and thiocyanate concentrations and for haematological studies; pancreas and thyroid gland were collected for histopathological study. The group receiving the highest dose of cyanide showed lower body weight gains and carcase weights and a decrease in plasma T3 concentrations compared to the control group. Reabsorption vacuoles in follicular colloid and normocytic normochromic anaemia were observed in the experimental animals. Inhibition of peripheral conversion of T4 to T3 is suggested. However, no diabetogenic effects were observed.

Lack of protective action of cysteine against the fetotoxic effect of monocrotaline.

Monocrotaline (MCT), a pyrrolizidine alkaloid present in Crotalaria species, has hepatotoxic, nephrotoxic, pneumotoxic and fetotoxic effects. However, the toxic effects of exposure to MCT in adult rats can be prevented by cysteine. Thus, the present study was conducted to evaluate the possible prevention by cysteine of the toxic effects of MCT on pregnant rats. Thirty-six pregnant rats were used. The females in the experimental groups were fed ration containing 0.02% MCT, 0.02% MCT + 1% cysteine, or 1% cysteine from day 6 to day 21 of pregnancy; the control group was fed only common ration for the same period of time. All rats were killed on day 21 of pregnancy and their blood was collected for determination of liver and kidney function. General toxicity to pregnant dams was assessed. Fetuses were removed by caesarian section and embryofetotoxic parameters were examined. Results showed impaired body weight gain in rats fed MCT, with or without cysteine supplementation. Plasma levels of AST, ALT, LDH, GGT, urea and creatinine were increased in MCT animals compared to controls. The pathology study revealed lesions only in dams from the MCT group. The weights of the placentas and fetuses of the MCT and MCT + cysteine groups were significantly lower than those of the control group. Thus, the present data suggests some protective action of 1% of cysteine in ration against the toxic effects of MCT on the dams but not on the litter.

Does prolonged cyanide exposure have a diabetogenic effect?

Cyanide exposure through cassava consumption has been associated with the development of malnutrition-related diabetes mellitus (MRDM). However, there are few experimental reproductions of this disease. In the present study 42 rats received 0, 9.0 or 12.0 mg KCN/kg bw/d for 15 d, 26 pigs were dosed with 0, 2.0, 4.0 or 6.0 mg KCN/kg for 74 d, and 34 goats received 0, 0.3, 0.6, 1.2 or 3.0 mg KCN/kg for 5 mo. At the end of each experimental period, plasma samples were obtained for glucose and thiocyanate measurement, and the pancreas was collected for histopathologic study. No significant differences in plasma glucose concentrations occurred between groups. The pancreas had no pathology. Chronic cyanide exposure did not promote diabetogenic effects in rats, swine or goats, suggesting that cyanide is not responsible for MRDM in humans.